Porcine TNF-alpha ELISA

    


RayBio® Porcine (pig) TNF alpha ELISA Kit for cell culture supernatants, plasma, and serum samples.



 ELP-TNFa-1

Antigen Information

Accession Number:
  • P23563

Gene ID:
  • 397086

Gene Symbols:
  • TNF

  • TNFA

  • TNFSF2

Protein Name & Synonyms:
Tumor necrosis factor (Cachectin) (TNF-alpha) (Tumor necrosis factor ligand superfamily member 2) (TNF-a) [Cleaved into: Tumor necrosis factor, membrane form (N-terminal fragment) (NTF) Intracellular domain 1 (ICD1) Intracellular domain 2 (ICD2) C-domain 1 C-domain 2 Tumor necrosis factor, soluble form]
Target Species:
  • Porcine (pig)

Assay Format

Detection Range:
20 pg/ml - 10 ng/ml
Recommended Dilution (serum & plasma):
2 fold
Sensitivity:
20 pg/ml

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Specificity:
The sandwich ELISA antibody pair detects Porcine TNF-alpha.
Number of Targets Detected:
1
Species Detected:
  • Porcine (pig)

Compatible Sample Types:
  • Cell Culture Supernatants

  • Plasma

  • Serum

Design Principle:
  • Sandwich-based

Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

Size:
1, 2, or 5 x 96-Well Strip Microplate Kit

Product Features

Additional Notes
  • Strip plates and additional reagents allow for use in multiple experiments

  • Quantitative protein detection

  • Establishes normal range

  • The best products for confirmation of antibody array data

Standard Curves

Porcine TNF-alpha ELISA(图1)

Porcine TNF-alpha ELISA(图2)

Spiking & Recovery Results

Porcine TNF-alpha ELISA(图3)

Linearity Results

Porcine TNF-alpha ELISA(图4)

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate

  • Wash Buffer

  • Stop Solution

  • Assay Diluent(s)

  • Lyophilized Standard

  • Biotinylated Detection Antibody

  • Streptavidin-Conjugated HRP

  • TMB One-Step Substrate

Other Materials Required
  • Distilled or deionized water

  • Precision pipettes to deliver 2 µl to 1 µl volumes

  • Adjustable 1-25 µl pipettes for reagent preparation

  • 100 µl and 1 liter graduated cylinders

  • Tubes to prepare standard and sample dilutions

  • Absorbent paper

  • Microplate reader capable of measuring absorbance at 450nm

  • Log-log graph paper or computer and software for ELISA data analysis

Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.

  2. Add 100 µl of standard or sample to each well.

  3. Incubate 2.5 h at RT or O/N at 4°C.

  4. Add 100 µl of prepared biotin antibody to each well.

  5. Incubate 1 h at RT.

  6. Add 100 µl of prepared Streptavidin solution to each well.

  7. Incubate 45 min at RT.

  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.

  9. Incubate 30 min at RT.

  10. Add 50 µl of Stop Solution to each well.

  11. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.